• The Plant Pathology Journal
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• v.16 no.2
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• pp.118-124
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• 2000

The coat protein (CP) gene of kyuri green mottle mosaic virus zucchini strain (KGMMV-Z) isolated from zucchini (Cucurbita pepo) in Chonfu, Korea in 1999 was sequenced by the reverse transcription and polymerase chain reaction with degenerate and generate primers originated from tobamoviruses. The degenerate primers were very effective in amplification of KGMMV-Z CP region. The KGMMV-Z CP gene consisted of 486 nucleotides and had the same nucleotide length compared with those of cucurbit-infecting tobamoviruses. KGMMV-Z CP gene shared 43.8, 44.2, and 44.4% nucleotide sequence similarity with the CP gene of cucumber green mottle mosaic virus watermelon strain (CGMMZ-W), CGMMV-KW1, and CGMMV-SH, respectively, whereas three CGMMV strains among themselves showed 98.6-99.6% nucleotide similarity. The deduced amino acids of KGMMV-Z CP gene were 161 amino acid residues with the molecular weight of 17,181 daltons. The first 24 codons of KGMMV-Z CP gene corresponded to the sequences of the N-terminal amino acid of the viral capsid protein. The amino acid sequences of KGMMV-Z CP had 45.3% similarity compared with those of three CGMMV strains. However, the amino acid sequences of CGMMV strains were identical. These results showed that two cucurbit-infecting tobamovirus members, KGMMV-Z and CGMMV were genetically distantly related.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.12
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• pp.1774-1783
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• 2015

Milk lysozyme is the ubiquitous enzyme in milk of mammals. In this study, the cDNA sequence of a new chicken-type (c-type) milk lysozyme gene (YML), was cloned from yak mammary gland tissue. A 444 bp open reading frames, which encodes 148 amino acids (16.54 kDa) with a signal peptide of 18 amino acids, was sequenced. Further analysis indicated that the nucleic acid and amino acid sequences identities between yak and cow milk lysozyme were 89.04% and 80.41%, respectively. Recombinant yak milk lysozyme (rYML) was produced by Escherichia coli BL21 and Pichia pastoris X33. The highest lysozyme activity was detected for heterologous protein rYML5 (M = 1,864.24 U/mg, SD = 25.75) which was expressed in P. pastoris with expression vector $pPICZ{\alpha}A$ and it clearly inhibited growth of Staphylococcus aureus. Result of the YML gene expression using quantitative polymerase chain reaction showed that the YML gene was up-regulated to maximum at 30 day postpartum, that is, comparatively high YML can be found in initial milk production. The phylogenetic tree indicated that the amino acid sequence was similar to cow kidney lysozyme, which implied that the YML may have diverged from a different ancestor gene such as cow mammary glands. In our study, we suggest that YML be a new c-type lysozyme expressed in yak mammary glands that plays a role as host immunity.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.17
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• pp.7943-7957
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• 2015

Background and Aims: Colorectal cancer is one of the leading causes of death in the world. The aim of this study was to investigate the growth-suppression potentiality of a crude saponin extract (CSENS) prepared from medicinal herb, Nigella sativa, on human colon cancer cells, HCT116. Materials and Methods: HCT116 cells were subjected to increasing doses of CSENS for 24, 48 and 72 h, and then harvested and assayed for cell viability by WST-1. Flow cytometry analyses, cell death detection ELISA, fluorescent stains (Hoechst 33342 and acridine orange/ethidium bromide), DNA laddering and comet assays were carried out to confirm the apoptogenic effects of CSENS. Luciferase reporter gene assays, quantitative reverse transcription-polymerase chain reaction and Western blot analyses were performed to assess the impact of CAERS and CFEZO on the expression levels of key regulatory proteins in HCT116 cells. Results: The results demonstrated that CSENS inhibited proliferation and induced apoptosis. Apoptosis was confirmed by flow cytometry analyses, while CSENS-treated cells exhibited morphological hallmarks of apoptosis including cell shrinkage, irregularity in cellular shape, cellular detachment and chromatin condensation. Biochemical signs of apoptosis, such as DNA degradation, were observed by comet assay and gel electrophoresis. The pro-apoptotic effect of CSENS was caspase-3-independent and associated with increase of the Bax/Bcl-2 ratio. CSENS treatment down-regulated transcriptional and DNA-binding activities of NF-${\kappa}B$ and AP-1 proteins, associated with down-regulation of their target oncogenes, c-Myc, cyclin D1 and survivin. On the other hand, CSENS up-regulated transcriptional and DNA-binding activities of Nrf2 and expression of cytoprotective genes. In addition, CSENS modulated the expression levels of ERK1/2 MAPK, p53 and p21. Conclusions: These findings suggest that CSENS may be a valuable agent for treatment of colon cancer.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.11
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• pp.6315-6319
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• 2013

To study the differentiated expression of the proto-oncogene Pokemon in nasopharyngeal carcinoma (NPC) cell lines and tissues, mRNA and protein expression levels of CNE1, CNE2, CNE3 and C666-1 were detected separately by reverse transcription polymerase chain reaction (RT-PCR), real-time PCR and Western-blotting. The immortalized nasopharyngeal epithelial cell line NP69 was used as a control. The Pokemon protein expression level in biopsy specimens from chronic rhinitis patients and undifferentiated non keratinizing NPC patients was determined by Western-blotting and arranged from high to low: C666-1>CNE1>CNE2> CNE3>NP69. The Pokemon mRNA expression level was also arranged from high to low: CNE1>CNE2>NP69>C666-1>CNE3. Pokemon expression of NP69 and C666-1 obviously varied from mRNA to protein. The Pokemon protein level of NPC biopsy specimens was obviously higher than in chronic rhinitis. The data suggest that high Pokemon protein expression is closely associated with undifferentiated non-keratinizing NPC and may provide useful information for NPC molecular target therapy.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.11
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• pp.6411-6413
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• 2013

Adenocarcinoma of esophagus (AE) is a complex disease, affected by a variety of genetic and environmental factors. Much evidence has shown that the MutY glycosylase homologue (MUTYH) plays a key role in the pathogenesis of many cancers. However, there have been no reports on influence on AE in the Han Chinese population. The objective of this study was to investigate this issue. A gene-based association study was conducted using three single nucleotide polymorphisms(SNPs) reported in previous studies. The three SNPs (rs3219463, rs3219472, rs3219489) were genotyped in 207 unrelated AE patients and 249 healthy controls in a case-control study using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). The results revealed that the genotype distribution of rs3219472 differed between the case and control groups (OR=1.66,95%CI=1.11-2.48, P=0.012), indicating that an association may exist between MUTYH and AE. These findings support a signifcant role for MUTYH in AE pathogenesis in the Han Chinese population.

• Journal of dental hygiene science
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• v.15 no.4
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• pp.488-494
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• 2015

A recent report showed that nuclear factor of activated T cell (NFATc) 1 is a member of the NFAT family and is strictly implicated osteoblast differentiation and bone formation. Furthermore, the precise expression and function of NFATc1 in periodontal tissue remains unclear. Therefore, the purpose of this study was to investigate the function of NFATc1 in osteoblastic differentiation, and the underlying mechanism regulating periodontal regeneration in human periodontal ligament cells (hPDLCs). NFATc1 messenger RNA (mRNA) and protein levels were accessed by reverse transcription-polymerase chain reaction (RT-PCR) and western blot assay, respectively. Cell proliferation determined using MTT assay. Differentiation was evaluated by alkaline phosphatase activity and formation of calcium nodule with alizarin red S staining. The mRNA expression of osteoblastic differentiation related genes were examined by RT-PCR. Marked upregulation of NFATc1 mRNA and protein was observed in cells grown in osteogenic medium (OS). NFATc1 transactivation was detected in hPDLCs that had been incubated in OS for 14 days. Treatment with $10{\mu}M$ cyclosporine A (CsA), a known calcineurin inhibitor, reduced the proliferation of hPDLCs, while $5{\mu}M$ CsA had no effect. Inhibition of the calcineurin/NFATc1 pathway by CsA, attenuated OS-induced osteoblastic differentiation in hPDLCs. In summary, this study demonstrates for the first time that NFATc1 plays a key role in osteoblastic differentiation of hPDLCs and activation of NFATc1 could provide a novel mechanism for periodontal bone regeneration.

• Parasites, Hosts and Diseases
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• v.35 no.4
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• pp.295-298
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• 1997

Genonlic DNA from Metagonimn vokogawci and Metagonimw Miyata type was amplified by polymerase chain reaction based on the random amplification of polymorphic DNA (RAPDI technique. Eight random 10-mer oligonucleotide primers (OPA-02, 5-TGCCGAGCTG-3; OPA-09, 5-GGGTAACGCC-3; OPA-17, 5-GTGATCGCAG-3; OPA-11, 5-CAATCGCCGT-3; OPA-13, 5-CAGCACCCAC-3; OPA-17. 5-GACCGCTrGT-3; OPA-19, 5-CAAACGTCGG-3; OPA-20, 5-GTTGCGATCC-3) WITH A G+C CONTENT FO 60-70% (Kit A. Operon Technologies Inc., California, USAI could produce distinguishable banding patterns between the two Metngonimus species. From the results of this study, it was suggested that Metcsonimus Miyata type has a different DNA sequence from M. WOkQgGUIGi. Key words: Metcgonimw vokognwai, MetnBonimw Miyata type, random amplification of polymorphic DNA (RAPD)

• Journal of Plant Biotechnology
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• v.34 no.2
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• pp.145-152
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• 2007

Lactoferrin is an 80-kDa iron-binding glycoprotein known to exert many biological activities, such as facilitating iron absorption and having antimicrobial and anti-inflammatory effects. Rice can be a useful target for edible food plants to introduce human lactoferrin, because it has lower allergenicity and is likely to be safer than microorganisms or transgenic animals. A cDNA fragment encoding human lactoferrin (HLF) driven by the maize polyubiquitin promoter, along with herbicide resistance gene (bar) driven by CaMV 35S promoter, was introduced into rice (Oryza sativa L. cv. Dong Jin) using the Agrobacterium -mediated transformation system. Putative transformants were initially selected on the medium containing bialaphos. The stable integration of the bar and HLF genes into transgenic rice plants was further confirmed through polymerase chain reaction (PCR) and Southern blot analyses. The expression of the full length HLF protein from various tissues such as grains and young leaves of transgenic rice was verified by Western blot analysis. Analysis of progeny also demonstrated that introduced genes were stably inherited to the next generation at the Mendelian fashion.

• Molecular & Cellular Toxicology
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• v.5 no.3
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• pp.230-236
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• 2009

Acrolein, a known toxin in cigarette smoke, is the most abundant electrophilic $\alpha$, $\beta$-unsaturated aldehyde to which humans are exposed in a variety of environmental pollutants, and is also product of lipid peroxidation. Increased unsaturated aldehyde levels and reduced antioxidant status plays a major role in the pathogenesis of various diseases such as diabetes, Alzheimer's and atherosclerosis. The findings reported here show that low concentrations of acrolein induce heme oxygenase-1 (HO-1) expression in RAW 264.7 macrophages. HO-1 induction by acrolein and signal pathways was measured using reverse transcription-polymerase chain reaction, Western blot and immunofluorescence staining analyses. Inhibition of extracellular signal-regulated kinase activity significantly attenuated the induction of HO-1 protein by acrolein, while suppression of Jun N-terminal kinase and p38 activity did not affect induction of HO-1 expression. Moreover, rottlerin, an inhibitor of protein kinase $\delta$, suppressed the upregulation of HO-1 protein production, possibly involving the interaction of NF-E2-related factor 2 (Nrf2), which has a key role as a HO-1 transcription factor. Acrolein elevated the nuclear translocation of Nrf2 in nuclear extraction. The results suggest that RAW 264.7 may protect against acrolein-mediated cellular damage via the upregulation of HO-1, which is an adaptive response to oxidative stress.

• The Journal of the Institute of Internet, Broadcasting and Communication
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• v.14 no.5
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• pp.27-33
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• 2014

As UCI (Universal Content Identifier) identification system becomes a national standard for content distribution, many digital contents are distributed through UCI identification system. However, problems associated with copyright infringement and copyright contracts occur due to lack of copyright information. ICN (Integrated Copyright Number), a code system developed for copyright management, includes copyright-related information for the purposes of usage settlement and accounting, illegal usage monitoring, and content filtering. ICN, however, has no information about distribution and content properties. Therefore, the linkage and integration between UCI and ICN can be a basis for the transparent and effective distribution of digital contents. In this paper, we propose a model for linking and integrating UCI and ICN, developing the efficiency in the value chain of each identified system by analyzing political factors and key technical elements between the two identification systems.